Northwest of Lake City and Live Oak near Fort Union, the 2,300 acre Holton Creek Concervation Area has 12 miles of sandy doubletrack. The blue-blazed route generally follows the Suwannee River through old growth bottomland forest with big cypress trees. Check out the Champion Cypress Tree and the Cypress Sinks.
To reach the mountain biking trailhead from Lake City, head west on US10 to the US90 exit. Turn right and then right again on the first road. Follow to the end and turn right on Knight Road. Turn left on SR249 and continue across the Suwannee River. Turn right at the second unpaved road to the Holton parking area.
Overall, Holton Creek Concervation Area is the 43rd most popular mountain bike trail of all 58 mountain biking rides in Florida.